Interactions between nanosized materials and the brain.

The current rapid development of nanotechnologies and engineered nanomaterials (ENM) will impact the society in a major fashion during the coming decades. This development also causes substantial safety concerns. Among the many promising applications of ENM, products that can be used for diagnosis and treatment of diseases, including conditions that affect the nervous system, are under development. ENM can pass the blood brain barrier (BBB) and accumulate within the brain. It seems that the nano-form rather than the bulk form of the chemicals pass the BBB, and that there is an inverse relationship between particle size and the ability to penetrate the BBB. Although translocation of ENM to the brain is possible during experimental conditions, the health relevance for real-life situations is far from clear. One major reason for this is that studies have been using nanoparticle concentrations that are far higher than the ones that can be expected during realistic exposures. However, very high exposure to the CNS can cause effects on neurotransmission, redox homeostasis and behavior. Available studies have been focusing on possible effects of the first generation of ENM. It will be necessary to study possible health effects also of expected novel sophisticated materials, independent of the outcome of present studies. The prospects for intended or targeted medical applications are promising since it has been shown that ENM can be made to pass the BBB and reach specific regions or cells within the brain.

Consumption of bee pollen affects rat ovarian functions.

The aim of this study was to examine possible effects of bee pollen added to the feed mixture (FM) on rat ovarian functions (secretion activity and apoptosis). We evaluated the bee pollen effect on the release of insulin-like growth factor I (IGF-I) and steroid hormones (progesterone and estradiol), as well as on the expression of markers of apoptosis (Bcl-2, Bax and caspase-3) in rat ovarian fragments. Female rats (n = 15) were fed during 90 days by FM without or with rape seed bee pollen in dose either 3 kg/1000 kg FM or 5 kg/1000 kg FM. Fragments of ovaries isolated from rats of each group (totally 72 pieces) were incubated for 24 h. Hormonal secretion into the culture medium was detected by RIA. The markers of apoptosis were evaluated by Western blotting. It was observed that IGF-I release by rat ovarian fragments was significantly (p < 0.05) decreased; on the other hand, progesterone and estradiol secretion was increased after bee pollen treatment at dose 5 kg/1000 kg FM but not at 3 kg/1000 FM. Accumulation of Bcl-2 was increased by bee pollen added at 3 kg/1000 kg FM, but not at higher dose. Accumulation of Bax was increased in ovaries of rats fed by bee pollen at doses either 3 or 5 kg/1000 kg FM, whilst accumulation of caspase-3 increased after feeding with bee pollen at dose 5 kg/1000 kg FM, but not at 3 kg/1000 kg FM. Our results contribute to new insights regarding the effect of bee pollen on both secretion activity (release of growth factor IGF-I and steroid hormones progesterone and estradiol) and apoptosis (anti- and pro-apoptotic markers Bcl-2, Bax and caspase-3). Bee pollen is shown to be a potent regulator of rat ovarian functions.

[Crural arteries PTA in patients with diabetes]. / PTA krurálnych tepien u diabetikov.

AIM: The authors analyse the patients hospitalized with a neuroischemic diabetic foot. They evaluate experiences and results of the crural arteries percutaneous transluminal angioplasty (PTA) in the diabetic patients. METHOD: It is a retrospective analysis of the patients hospitalized during years 2005-2008 at the Department of Vascular Surgery of the Teaching Hospital J. A. Reiman Presov with the diabetic foot. During the study period 265 hospitalization in 205 patients were registered for complications of the diabetic foot. PTA of the crural arteries was performed on 74 lower extremities in 68 patients. 133 crural arteries were revascularized in this group of the patients. Indication for execution of the crural arteries PTA in the diabetic patients was critical limb ischemia on 68 revascularized lower extremities (91.9%). RESULTS: Technical success (residual stenosis no more than 30%, unobstructed revascularized crural artery to periphery) was achieved on 93 revascularized crural arteries (69.92%). They have control examination minimally 12 months after PTA in 54 revascularized lower extremities of the 68 patients with the neuroischemic diabetic foot. From this subset of the patients 28 lower extremities (51.85%) are healed according to clinical examination, on 7 lower extremities (12.96%) clinical signs of critical limb ischemia remain, 10 times (18.52%) low amputation was performed and 9 times (16.67%) high amputation was executed. CONCLUSION: The authors believe that PTA of the crural arteries is safe and effective method of critically ischemic diabetic foot revascularization. If revascularization is successful, it can effectively contributes to the lower extremity salvage of the patient with neuroischemic diabetic foot.

[Treatment of thrombotic complications of surgical infrainguinal revascularizations]. / Liecba trombotických komplikácií chirurgických infrainguinálnych revaskularizacných výkonov.

AIM: Authors in the work analyse the patients hospitalized with thrombosis of the surgical infrainguinal reconstruction. They evaluate the causes of reconstructions occlusions, therapeutic options in the treatment and their experiences with treatment of these patients. METHOD: It is the retrospective analysis of the patients hospitalized during years 2001-2006 at the Department of Vascular Surgery of the Teaching Hospital J. A. Reimana in Presov with thrombosis of the surgical infrainguinal reconstruction. During the study period 723 surgical infrainguinal revascularization procedures were performed. The basic therapeutic approach in the care of these patients is anticoagulation and digital subtraction angiography, then they decide between percutaneous endovascular treatment, surgical thrombectomy, reoperation with implantation of the new reconstruction or conservative treatment. In the phase of irreversible ischemia high amputation is indicated. RESULTS: Thrombosis of the reconstruction was occured in 109 subjects (15.8%). Prothesis was thrombosed in 71 subjects (65.14%), vein in 38 subjects (34.86%). Percutaneous endovascular treatment was indicated in 3 subjects (2.8%), surgical thrombectomy with correction of the stenosis in 23 subjects (21.1%), reoperation with implantation of the new reconstruction in 53 subjects (48.6%),conservative approach in 21 subjects (19.3%), high amputation in 9 subjects (8.2%). The secondary patency after 12 months according to the life table method analysis after percutaneous endovascular treatment was 75%, after surgical thrombectomy of the graft with correction of stenosis 37% and after reoperation with implantation of the new reconstruction 72%. CONCLUSION: Authors believe that the best treatment in the care of the patients with thrombosis of the surgical infrainguinal reconstruction and critical limb ischemia is implantation of the new secondary reconstruction with autologous vein.

MRI-based brain volumetry in chronic whiplash patients: no evidence for traumatic brain injury.

INTRODUCTION: Cognitive complaints, such as poor concentration and memory deficits, are frequent after whiplash injury and play an important role in disability. The origin of these complaints is discussed controversially. Some authors postulate brain lesions as a consequence of whiplash injuries. Potential diffuse axonal injury (DAI) with subsequent atrophy of the brain and ventricular expansion is of particular interest as focal brain lesions have not been documented so far in whiplash injury. OBJECTIVE: To investigate whether traumatic brain injury can be identified using a magnetic resonance (MR)-based quantitative analysis of normalized ventricle-brain ratios (VBR) in chronic whiplash patients with subjective cognitive impairment that cannot be objectively confirmed by neuropsychological testing. MATERIALS AND METHODS: MR examination was performed in 21 patients with whiplash injury and symptom persistence for 9 months on average and in 18 matched healthy controls. Conventional MR imaging (MRI) was used to assess the volumes of grey and white matter and of ventricles. The normalized VBR was calculated. RESULTS: The values of normalized VBR did not differ in whiplash patients when compared with that in healthy controls (F = 0.216, P = 0.645). CONCLUSIONS: This study does not support loss of brain tissue following whiplash injury as measured by VBR. On this basis, traumatic brain injury with subsequent DAI does not seem to be the underlying mechanism for persistent concentration and memory deficits that are subjectively reported but not objectively verifiable as neuropsychological deficits.

[The isolated internal iliac artery aneurysms]. / Izolované aneuryzmy arteria iliaca interna.

The isolated iliac artery aneurysm is a rare disease. Diagnosis and treatment of these aneurysms according to their anatomic location is difficult. Authors describe diagnostic-therapeutic approach in the care of three patients at the vascular unit of the Surgical Clinic of the Teaching Hospital in Presov. They analyze different methods of treatment with emphasis on endovascular therapeutic methods mainly in the therapy of polymorbid high risk patients.

Comparative study of cell cycle kinetics and induction of apoptosis or necrosis after exposure of human Mono Mac 6 cells to radiofrequency radiation.

The possible harmful effects of radiofrequency electromagnetic fields (RF EMFs) are controversial. We have used human Mono Mac 6 cells to investigate the influence of RF EMFs in vitro on cell cycle alterations and BrdU uptake, as well as the induction of apoptosis and necrosis in human Mono Mac 6 cells, using flow cytometry after exposure to a 1,800 MHz, 2 W/kg specific absorption rate (SAR), GSM-DTX signal for 12 h. No statistically significant differences in the induction of apoptosis or necrosis, cell cycle kinetics, or BrdU uptake were detected after RF EMF exposure compared to sham or incubator controls. However, in the positive control cells treated with gliotoxin and PMA (phorbol 12 myristate-13 acetate), a significant increase in apoptotic and necrotic cells was seen. Cell cycle analysis or BrdU incorporation for 72 h showed no differences between RF EMF- or sham-exposed cells, whereas PMA treatment induced a significant accumulation of cells in G(0)/G(1)-phase and a reduction in S-phase cells. RF EMF radiation did not induce cell cycle alterations or changes in BrdU incorporation or induce apoptosis and necrosis in Mono Mac 6 cells under the exposure conditions used.

ROS release and Hsp70 expression after exposure to 1,800 MHz radiofrequency electromagnetic fields in primary human monocytes and lymphocytes.

The aim of this study is to investigate if 1,800 MHz radiofrequency electromagnetic fields (RF-EMF) can induce reactive oxygen species (ROS) release and/or changes in heat shock protein 70 (Hsp70) expression in human blood cells, using different exposure and co-exposure conditions. Human umbilical cord blood-derived monocytes and lymphocytes were used to examine ROS release after exposure to continuous wave or different GSM signals (GSM-DTX and GSM-Talk) at 2 W/kg for 30 or 45 min of continuous or intermittent (5 min ON/5 min OFF) exposure. The cells were exposed to incubator conditions, to sham, to RF-EMF, or to chemicals in parallel. Cell stimulation with the phorbol ester phorbol-12-myristate-13-acetate (PMA; 1 microM) was used as positive control for ROS release. To investigate the effects on Hsp70 expression, the human monocytes were exposed to the GSM-DTX signal at 2 W/kg for 45 min, or to heat treatment (42 degrees C) as positive control. ROS production and Hsp70 expression were determined by flow cytometric analysis. The data were compared to sham and/or to control values and the statistical analysis was performed by the Student's t-test (P<0.05). The PMA treatment induced a significant increase in ROS production in human monocytes and lymphocytes when the data were compared to sham or to incubator controls. After continuous or intermittent GSM-DTX signal exposure (2 W/kg), a significantly different ROS production was detected in human monocytes if the data were compared to sham. However, this significant difference appeared due to the lowered value of ROS release during sham exposure. In human lymphocytes, no differences could be detected if data were compared either to sham or to incubator control. The Hsp70 expression level after 0, 1, and 2 h post-exposure to GSM-DTX signal at 2 W/kg for 1 h did not show any differences compared to the incubator or to sham control.

Free radical release and HSP70 expression in two human immune-relevant cell lines after exposure to 1800 MHz radiofrequency radiation.

The goal of this study was to investigate whether radiofrequency (RF) electromagnetic-field (EMF) exposure at 1800 MHz causes production of free radicals and/or expression of heat-shock proteins (HSP70) in human immune-relevant cell systems. Human Mono Mac 6 and K562 cells were used to examine free radical release after exposure to incubator control, sham, RF EMFs, PMA, LPS, heat (40 degrees C) or co-exposure conditions. Several signals were used: continuous-wave, several typical modulations of the Global System for Mobile Communications (GSM): GSM-non DTX (speaking only), GSM-DTX (hearing only), GSM-Talk (34% speaking and 66% hearing) at specific absorption rates (SARs) of 0.5, 1.0, 1.5 and 2.0 W/kg. Heat and PMA treatment induced a significant increase in superoxide radical anions and in ROS production in the Mono Mac 6 cells when compared to sham and/or incubator conditions. No significant differences in free radical production were detected after RF EMF exposure or in the respective controls, and no additional effects on superoxide radical anion production were detected after co-exposure to RF EMFs+PMA or RF EMFs+LPS. The GSM-DTX signal at 2 W/kg produced a significant difference in free radical production when the data were compared to sham because of the decreasing sham value. This difference disappeared when data were compared to the incubator controls. To determine the involvement of heat-shock proteins as a possible inhibitor of free radical production, we investigated the HSP70 expression level after different RF EMF exposures; no significant effects were detected.

Hsp70 expression and free radical release after exposure to non-thermal radio-frequency electromagnetic fields and ultrafine particles in human Mono Mac 6 cells.

The contemporary urban environment has become increasingly complex in its composition, leading to discussions regarding possible novel health effects. Two factors that recently have received considerable attention are ultrafine particles (UFP; <0.1 microm) produced by combustion processes and emissions from wireless communication devices like mobile phones that emit in the radio-frequency (RF) part of the spectrum. Several studies have shown biological effects of both these exposures in various cell systems. Here we investigate if exposure to UFP (12-14 nm, 100 microg/ml) and RF-electromagnetic fields (EMF; 2 W/kg specific absorption rate (SAR); continuous wave (CW) or modulated (217Hz or GSM-nonDTX)), alone or in combination influences levels of the superoxide radical anion or the stress protein heat-shock protein (Hsp70) in the human monocyte cell line Mono Mac 6. Heat treatment (42-43 degrees C, 1h) was used as positive control for both stress reaction and for heat development in the RF exposure setup. Our results clearly show that Mono Mac 6 cells are capable to internalise UFP, and that this phagocytic activity is connected to an increased release of free radicals. This increase (40-45% above negative control) is stronger than the effect of heat treatment. On the other hand, none of the employed RF exposures showed any effects on free radical levels. Co-exposure of RF and UFP did not potentiate the UFP effect either. Our investigations showed a significantly increased Hsp70 expression level by heat treatment in a time-dependent manner, whereas UFP, RF, or UFP+RF were without any effect. Therefore, we conclude that in the investigated Mono Mac 6 cells, RF exposure alone or in combination with UFP cannot influence stress-related responses.

Short communication: hydroperoxides in circulating lipids from dairy cows: implications for bioactivity of endogenous-oxidized lipids.

This study was conducted to investigate the potential for increased oxidative stress of high- vs. average-producing dairy cows. Two experiments were performed using 11 and 13 Holstein cows (53 +/- 2 d postpartum). Lipohydroperoxides (LHP) were determined in serum lipids (experiment 1) and low-density lipoprotein (experiment 2) via oxidation of ferrous to ferric ions through LHP using thiocyanate as chromogen. In experiment 1, differing milk yield and milk energy output corresponded to different concentrations of LHP. In experiment 2, analysis of regression resulted in a significant relationship between milk yield and LHP. Phospholipids isolated from lipids with 6.5 microM of LHP evoked in monocytic cells a transient increase in superoxide formation, indicating inflammatory potential. The results show that high milk productivity can associate with oxidative stress indicated by oxidative modifications of circulating lipids and their changed bioactivity.

Modifications in cell cycle kinetics and in expression of G1 phase-regulating proteins in human amniotic cells after exposure to electromagnetic fields and ionizing radiation.

Low-frequency electromagnetic fields are suspected of being involved in carcinogenesis, particularly in processes that could be related to cancer promotion. Because development of cancer is associated with deregulated cell growth and we previously observed a magnetic field-induced decrease in DNA synthesis [Lange et al. (2002) Alterations in the cell cycle and in the protein level of cyclin D1p, 21CIP1, and p16INK4a after exposure to 50 HZ. MF in human cells. Radiat. Environ. Biophys.41, 131], this study aims to document the influence of 50 Hz, 1 mT magnetic fields (MF), with or without initial gamma-ionizing radiation (IR), on the following cell proliferation-relevant parameters in human amniotic fluid cells (AFC): cell cycle distribution, expression of the G1 phase-regulating proteins Cdk4, cyclin D1, p21CIP1 and p16INK4a, and Cdk4 activity. While IR induced a G1 delay and a dose-dependent G2 arrest, no discernible changes in cell cycle kinetics were observed due to MF exposure. However, a significant decrease in the protein expression of cyclin D1 and an increase in p21CIP1- and p16INK4a-expression could be detected after exposure to MF alone. IR-exposure caused an augmentation of p21CIP1- and p16INK4a- levels as well, but did not alter cyclin D1 expression. A slight diminution of Cdk4 activity was noticed after MF exposure only, indicating that Cdk4 appears not to act as a mediator of MF- or IR-induced changes in the cell cycle of AFC cells. Co-exposure to MF/IR affected neither cell cycle distribution nor protein expression or kinase activity additionally or synergistically, and therefore MF seems not to modify the mutagenic potency of IR.

[Changes in serum levels of lipid peroxidation products during labor and in the puerperium]. / Zmeny sérových hladín produktov lipoperoxidácie pocas pôrodu a v sestonedelí.

OBJECTIVE: To determine the activity of reactive oxygen species (ROS) during labour by characterizing changes in maternal serum levels of lipid peroxidation end-products--MDA and lipofuscin during labour and the early post-partum period. We also tried to evaluate the relationship between levels of lipid peroxides and some clinical characteristics of labour. DESIGN: Prospective study. SETTING: Department of Obstetrics and Gynaecology LFUK, Bratislava; Department of clinical laboratories, Ministry of defense SR, Bratislava. METHODS: Blood samples were obtained from 66 pregnant women with uncomplicated pregnancy at the end of labour and during the early post-partum period. The control group consisted of 19 pregnant women delivering by primary Cesarean section. Blood samples were examined for MDA and lipofuscin by HPLC method. We used paired and unpaired Student's t-test to statistically evaluate our results. RESULTS: MDA and lipofuscin levels in pregnant women delivering spontaneously compared to those delivering by C-section were significantly elevated (P < 0.05). MDA and lipofuscin levels in pregnant women during spontaneous labour or during by C-section compared to the levels in early post-partum period were not significantly increased. We have not found any correlation between the length of the labour and lipoperoxides concentration.

An entirely suprasellar Rathke's cleft cyst: case report and review of the literature.

Rathke's cleft cysts usually present with an intrasellar or a combined intrasellar and suprasellar localization. In this report we present a 58-year-old man with a Rathke's cleft cysts in an entirely suprasellar location with intact diaphragm and intact sella. Although this presentation of a Rathke's cleft cyst is extremely rare, it belongs in the differential diagnosis of entirely suprasellar lesions. The relevant literature is discussed.

Stimulation of phagocytosis and free radical production in murine macrophages by 50 Hz electromagnetic fields.

Effects of 50 Hz electromagnetic fields on phagocytosis and free radical production were examined in mouse bone marrow-derived macrophages. Macrophages were in vitro exposed to electromagnetic fields using different magnetic field densities (0.5-1.5 mT). Short-time exposure (45 min) to electromagnetic fields resulted in significantly increased phagocytic uptake (36.3% +/- 15.1%) as quantified by measuring the internalization rate of latex beads. Stimulation with 1 nM 12-0-tetradecanoylphorbol-13-acetate (TPA) showed the same increased phagocytic activity as 1 mT electromagnetic fields. However, co-exposure to electromagnetic fields and TPA showed no further increase of bead uptake, and therefore we concluded that because of the absence of additive effects, the electromagnetic fields-induced stimulation of mouse bone marrow-derived macrophages does not involve the protein kinase C signal transduction pathway. Furthermore, a significant increased superoxide production after exposure to electromagnetic fields was detected.

Micronucleus induction in Syrian hamster embryo cells following exposure to 50 Hz magnetic fields, benzo(a)pyrene, and TPA in vitro.

Electromagnetic fields (EMFs) have been associated with increased incidence of cancer suggested by epidemiological studies. To test the carcinogenic potency of EMF, the in vitro micronucleus assay with SHE cells has been used as a screening method for genotoxicity. A 50Hz magnetic field (MF) of 1mT field strength was applied either alone or with the tumour initiator benzo(a)pyrene (BP) or the tumour promoter 12-O-tetradecanoylphorbol-13-acetate (TPA). All three treatments were applied in single, double or triple treatment regimes. MF or TPA (1nM) alone did not affect the number of micronuclei (MN) in initiated and non-initiated SHE cells. Changing the schedule of the typical initiation protocol, namely applying the initiator (BP) during exposure to MF, results in an 1.8-fold increased MN formation compared to BP treatment alone. Combined experiment with BP, TPA and MF did not cause further MN formation. Since initiation during MF exposure caused a significant increased MN formation, our findings suggest that MFs enhance the initiation process of BP. We think that this MF-enhanced co-carcinogenic effect is caused by an indirect "cell activation" process. The resulting genomic instability is proposed to be due to free radicals and/or to the unscheduled "switching-on" of signal transduction pathways.

Apoptosis induction and micronucleus formation after exposure to the Auger electron emitter zinc-65 in a human cell line.

Induction of apoptosis and micronucleus formation has been studied in a transformed human squamous cell carcinoma cell line (SCL-II) after exposure to the Auger electron emitter Zinc-65 (65Zn) and after external low-LET radiation. Exposure to non-radioactive Zn and unirradiated cells served as controls. Studies on the cellular uptake of 65Zn2+ have been carried out in vitro and conventional dosimetric models have been applied to derive the absorbed radiation dose. Auger electrons, generated during decay of 65Zn2+, are strong inducers of micronuclei as well as of apoptosis, in comparison to external low-LET irradiation. The relative biological effectiveness has been determined and was found to be in the range of 1.2-2.7 for the two investigated biological endpoints, depending on which mathematical model for describing the dose-effect curves was used. A non-uniform distribution of intracellular Zn2 + was observed, showing a strong signal in the perinuclear region. We conclude that separate radiation weighting factors for Auger electrons should be established depending on the nuclide and its ability to interact with the DNA (e.g. 65Zn by Zinc-finger proteins).

Absence of synergistic effects on micronucleus formation after exposure to electromagnetic fields and asbestos fibers in vitro.

Exposure of human amniotic fluid (AFC) cells to horizontally applied magnetic fields (hMF) of 50 Hz and 1 mT generated in a Helmholtz-coil system leads to a significant increase in micronucleus frequency (MN), without affecting cell proliferation. To investigate whether hMF-exposure has an additive or synergistic effect on the genotoxic capacity of asbestos fibers, MN induction was investigated in hMF pre-exposed cells, treated before or after with asbestos (1 microg/cm2). Neither synergistic nor additive effects on MN induction were observed. The results indicate, that under our experimental conditions, exposure to hMF and treatment with asbestos fibers possess genotoxic capability, but no interactive effects, in AFC cells.

Delayed cytotoxic and genotoxic effects in a human cell line following X-irradiation.

BACKGROUND: In order to clarify the relationship between delayed reproductive death and radiation-induced genomic instability, the colony-forming efficiency of surviving, irradiated human squamous carcinoma cells and centromere positive as well as centromere negative micronuclei in surviving progeny were examined. MATERIALS AND METHODS: Colony-forming ability and micronucleus (MN) frequency in binucleated cells 24 h after the addition of cytochalasin B during 2 weeks of post-irradiation growth were determined in a squamous cell carcinoma cell line (SCL-II) of human origin. In addition, centromeres in micronuclei were detected using FISH. RESULTS: In the human epithelial cell line used for these experiments, delayed reproductive death was pronounced and persisted for at least 2 weeks after irradiation. Although there is evidence for an increased rate of centromere positive micronuclei, but not of centromere negative micronuclei, arising during the first week of post-irradiation proliferation, this decreases later while the rate of delayed reproductive death remains elevated. CONCLUSION: In the studied cell line, the observed delayed reproductive death is not closely related to the investigated criteria of radiation-induced genomic instability. This casts doubt on the common assumption that delayed reproductive death is a direct manifestation of radiation-induced genomic instability.

Micronucleus formation in human amnion cells after exposure to 50 Hz MF applied horizontally and vertically.

Micronucleus (MN) induction as a genotoxic effect of extremely-low-frequency electromagnetic fields (ELF-EMF, 50 Hz, 1 mT) was studied in human amniotic fluid cells (AFC) after continuous exposure to magnetic fields (MF), oriented horizontally and vertically with respect to the surface of the culture medium, at different time points. To compare the effectiveness of different exposure systems, a Helmholtz-coil system and a so-called Merritt-coil system was used. A statistically significant increase in MN frequency could be detected in exposed cells compared to controls after 72 h continuous exposure to MF applied vertically in the Merritt-coil system, while no effect was found after exposure in the Helmholtz-coil system. Furthermore, a significant increase in MN induction occurred after 24, 48 and 72 h exposure to MF applied horizontally in the Helmholtz-coil system in comparison to controls, whereas horizontally MF generated in the Merritt-coil system induced no genotoxic effects. To exclude suppression of indirect EMF-induced DNA-lesions, we studied MN formation in the presence of N-Acetyl-p-aminophenol (APAP, Paracetamol(R)), which is an inhibitor of DNA-repair mechanisms. We found a dose-dependent increase of MN formation in APAP-treated AFC cells, but no significant further increase in MN frequency after additional MF exposure. Therefore we conclude, that EMF-induced MN formation is not caused by directly or indirectly induced clastogenic mechanisms. The obtained results show that the orientation of MF with respect to the cell culture dish and the physical condition of the exposure system is of major importance for the induction of micronuclei in certain cell types. Therefore, the reason for inconsistent results published in the literature may be caused by the variability of exposure systems, the exposure conditions and the cell types used.